Development of programmable small DNA-binding molecules with epigenetic activity for induction of core pluripotency genes

Epigenetic modifications that govern the gene expression are often overlooked with the design of artificial genetic switches. N-Methylpyrrole-N-methylimidazole (PI) hairpin polyamides are programmable small DNA binding molecules that have been studied in the context of gene regulation. Recently, we synthesized a library of compounds by conjugating PI polyamides with SAHA, a chromatin-modifier. Among these novel compounds, PI polyamide-SAHA conjugate 1 was shown to epigenetically activate pluripotency genes in mouse embryonic fibroblasts. Here, we report the synthesis of the derivatives of conjugate 1 and demonstrate that these epigenetically active molecules could be developed to improve the induction of pluripotency factors.     

Value of a newly sequenced bacterial genome

Next-generation sequencing(NGS) technologies have made high-throughput sequencing available to medium- and small-size laboratories, culminating in a tidal wave of genomic information. The quantity of sequenced bacterial genomes has not only brought excitement to the field of genomics but also heightened expectations that NGS would boost antibacterial discovery and vaccine development. Although many possible drug and vaccine targets have been discovered, the success rate of genome-based analysis has remained below expectations. Furthermore, NGS has had consequences for genome quality, resulting in an exponential increase in draft(partial data) genome deposits in public databases. If no further interests are expressed for a particular bacterial genome, it is more likely that the sequencing of its genome will be limited to a draft stage, and the painstaking tasks of completing the sequencing of its genome and annotation will not be undertaken. It is important to know what is lost when we settle for a draft genome and to determine the "scientific value" of a newly sequenced genome. This review addresses the expected impact of newly sequenced genomes on antibacterial discovery and vaccinology. Also, it discusses the factors that could be leading to the increase in the number of draft deposits and the consequent loss of relevant biological information.     

Safety and toxicological evaluation of Rhizopora mucronata (a mangrove from Vellar estuary,India): assessment of mutagenicity,genotoxicity and in vivo acute toxicity

The present was carried out to evaluate the toxicity of methanolic leaf extract of Rhizophora mucronata (MERM) under in vivo and in vitro conditions. Mutagenicity of MERM (up to 4,000 μg/plate) evaluated by Salmonella/microsome assay (TA98, TA100, TA1535 and TA1538 strains), with or without metabolic activation showed no mutagenic effect in any of the tester strain. Evaluation of genotoxicity (comet assay) and cytotoxicity in PBMC revealed that MERM showed no significant difference in comet tail moment (TM) and tail scores and cytotoxicity up to 24 h respectively. In acute toxicity studies, oral administration of single doses of MERM (250–2,000 mg/kg) in Wistar rats produced neither mortality nor any noticeable changes in behavior. Hematological and biochemical parameters showed no difference, except for a significant increase in ALT and AST at the highest dose. Histopathological findings revealed hepatotoxicity and neurotoxicity at highest dose of extract. In subacute toxicity studies administration of MERM (1,000 mg/kg) for 28 consecutive days neither altered the body weight gain nor behavioral parameters. No significant change was observed in the hematological and biochemical parameters analyzed. Histopathological examination showed normal architecture suggesting no morphological disturbances. Collectively, these data demonstrate that consumption of MERM for various medicinal purpose is safe.     

Purification, immobilization, and characterization of nattokinase on PHB nanoparticles

In this study, nattokinase was purified from Bacillus subtilis using ion exchange chromatography and immobilized upon polyhydroxybutyrate (PHB) nanoparticles. A novel strain isolated from industrial dairy waste was found to synthesize polyhydroxyalkanoates (PHA) and the strain was identified as Brevibacterium casei SRKP2. PHA granules were extracted from 48 h culture and the FT-IR analysis characterized them as PHB, a natural biopolymer from B. casei. Nanoprecipitation by solvent displacement technique was used to synthesize PHB nanoparticles. PHB nanoparticles were characterized using transmission electron microscopy and particle size ranged from 100–125 nm. Immobilization of nattokinase upon PHB nanoparticles resulted in a 20% increase in the enzyme activity. Immobilization also contributed to the enhanced stability of the enzyme. Moreover, the activity was completely retained on storage at 4 °C for 25 days. The method has proven to be highly simple and can be implemented to other enzymes also.     

Labeling of bovine corpus luteal plasma membrane human chorionic gonadotropin or luteinizing hormone (hCG/LH) receptor and its purification and properties.

A method has been developed for labeling receptors for human chorionic gonadotropin or luteinizing hormone ($\text{hCG}\text{LH}$) present on bovine corpus luteal plasma membranes. It consists of four steps: (a) protection of the receptor by treating the plasma membranes with hCG; (b) iodination of the membranes with KI using glucose, glucose oxidase, and lactoperoxidase; (c) unmasking the receptor with either 2 m NaCl, 1 m guanidine hydrochloride, or rabbit anti-hCG; and (d) reiodination of the membranes using Na¹³¹I. After solubilization by successive treatments with Sepharose-concanavalin A and Sepharose-hCG and finally by preparative disc electrophoresis, the resulting purified receptor after electrophoresis in polyacrylamide gel showed a single radioactive band containing receptor activity. This highly purified receptor is fairly stable and retains its hormonal specificity, binding affinity, and pH optimum. It was observed that the receptor alone or as a complex with the hormone tends to aggregate. The receptorhormone complex does not dissociate during polyacrylamide-gel electrophoresis.     

Energy extraction efficiency in some chosen insects

Summary The termsenergy extraction efficiency andabsorption/assimilation efficiency are defined. The grasshopperPoecilocerus pictus extracts 15 or 27% energy contained in the food plantCalotropis gigantea, when the test animals were fed from hatching to death at 26 or 36°C. The energy extraction efficiency of some insects (Bombyx andHyalophora: terrestrial herbovores;Pyrrhosoma: aquatic carnivore) is about 7%, though their assimilation efficiency ranges from 42 to 97%. High ash content of food organisms deviates the energy extraction efficiency inPyrrhosoma and the aquatic dipteran herbivoreHedriodiscus.     

Molecular cloning of growth hormone encoding cDNA of Indian major carps by a modified rapid amplification of cDNA ends strategy

A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences ofLabeo rohita, Cirrhina mrigala andCatla catla have been cloned, characterized and overexpressed inEscherichia coli. These sequences show 96–98% homology to each other and are about 85% homologous to that of common carp. Besides, an attempt has been made for the first time to describe a 3-D model of the fish GH protein.     

A new toxin isolated from Xanthomonas malvacearum which inhibits mitochondrial ATP--ADP translocase

The pathogenic effect of cotton blight disease is influenced by Xanthomonas malvacearum toxin. The toxin has been highly purified and the interaction between the toxin and the energy-generating system of mitochondria has been characterized. The results show that the toxin inhibits the ATP-ADP translocase system of the mitochondria.     

Optimization of anticancer exopolysaccharide production from probiotic Lactobacillus acidophilus by response surface methodology

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths in the Western world. Recently, much attention has been focused on decreasing the risk of CRC by consuming probiotics. In the present study, exopolysaccharide (EPS) extracted from Lactobacillus acidophilus was found to inhibit the growth of CaCo2 colon cancer cell line in a dose-dependent manner. The experiment was performed in both normoxic and hypoxic conditions, and EPS was found to reduce the survival of CaCo2 cell line in both the conditions. Quantitative polymerase chain reaction (qPCR) studies demonstrated that EPS treatment upregulated the expression of peroxisome proliferator activator receptor-γ (PPAR-γ) in both normoxia and hypoxia conditions, whereas it upregulated the expression of erythropoietin (EPO) in the normoxic condition, but there was no significant expression under hypoxic conditions. Hence, the EPS production was optimized by Plackett–Burman design followed by central composite rotatory design. The optimized production of EPS at 24 hr was found to be 400 mg/L. During batch cultivation the production peaked at 21 hr, resulting in an EPS concentration of 597 mg/L.     

Yolk energy utilization in Macrobrachium nobilii (Henderson & Mathai)

Energy content of Macrobrachium nobilii (Henderson & Mathai) is 0.19 cal in a freshly laid egg, 0.14 cal in a freshly hatched zoea, and 0.09 cal in an 8-day-old non-feeding zoea. These values indicate that the yolk is utilized with an efficiency of 74%, and that yolk equivalent to 0.1 cal is provided in an egg to meet embryonic and larval metabolism. The high yolk utilization efficiency and the high calorific content of the freshly hatched zoeae (5753 cal/g dry wt) are considered as adaptations to larval migration.